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Determination method of TritonX-100 residual quantity in antitoxin/ antiserum

A measurement method and anti-toxin technology, applied in the biological field, can solve problems such as effective control, achieve the effect of accurate method, reduce loss, and ensure accuracy

Inactive Publication Date: 2012-07-18
玉溪九洲生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Since there is no general method for the determination of Triton X-100 residues in antitoxin and antiserum products, it is difficult to effectively control the quality of antitoxin and antiserum products that have been inactivated by organic solvents and detergents

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Determination of Triton X-100 residues in tetanus antitoxin:

[0024] Instruments: 1. Agilent 1200 high performance liquid chromatography and ChemStations workstation

[0025] 2. Mettler AE 240 electronic balance

[0026] Reagents and reagents: Triton X-100 reference substance (purchased from Sigma, batch number: 118K0095); methanol was chromatographically pure (Fisher Chemical Reagent Company, USA); water was ultrapure water; other reagents were analytically pure.

[0027] Go through the following steps:

[0028] (1) Dissolve the reference substance Triton X-100 in water to make a reference solution containing 10 μg of Triton X-100 per 1 mL of solution;

[0029] (2) Take 0.5mL of tetanus antitoxin, the test product inactivated by the conventional organic solvent / detergent method, pass through the pre-activated solid phase extraction column, wash with water first, and then use 5% volume concentration Wash with methanol solution, then elute with 1mL of methanol, and c...

Embodiment 2

[0032] Determination of Triton X-100 residues in anti-rabies serum:

[0033] Instruments: 1. Agilent 1200 high performance liquid chromatography and ChemStations workstation

[0034] 2. Mettler AE 240 electronic balance

[0035] Reagents and reagents: Triton X-100 reference substance (purchased from Sigma, batch number: 118K0095); methanol was chromatographically pure (Fisher Chemical Reagent Company, USA); water was ultrapure water; other reagents were analytically pure.

[0036] Go through the following steps:

[0037] (1) Dissolve the reference substance Triton X-100 in water to make a Triton X-100 reference solution containing 5 μg per 1 mL;

[0038] (2) Take 1mL of anti-rabies serum, which has been inactivated by the conventional organic solvent / detergent method, pass through a pre-activated solid-phase extraction column, wash with water, and then use methanol with a volume concentration of 7%. Solution cleaning, then eluting with 1mL of methanol, collecting the eluent...

Embodiment 3

[0041] Determination of Triton X-100 residues in antivenom:

[0042]Instruments: 1. Agilent 1200 high performance liquid chromatography and ChemStations workstation

[0043] 2. Mettler AE 240 electronic balance

[0044] Reagents and reagents: Triton X-100 reference substance (purchased from Sigma, batch number: 118K0095); methanol was chromatographically pure (Fisher Chemical Reagent Company, USA); water was ultrapure water; other reagents were analytically pure.

[0045] Go through the following steps:

[0046] (1) Dissolve the reference substance Triton X-100 in water to make a Triton X-100 reference solution containing 20 μg per 1 mL;

[0047] (2) Take 0.5 mL of antivenom serum, which has been inactivated by the conventional organic solvent / detergent method, pass through the pre-activated solid phase extraction column, wash with water first, and then use 10% volume concentration Wash with methanol solution, then elute with 1mL of methanol, and collect the eluent to be th...

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Abstract

The invention provides a determination method of TritonX-100 residual quantity in antitoxin / antiserum, which comprises the following steps: respectively adsorbing comparison product solution and test sample solution; measuring by using a high-efficient liquid chromatography method; and carrying out conventional calculation according to the ratio of a peak area to solution concentration to obtain the TritonX-100 residual quantity in antitoxin / antiserum. With the method, macromolecular substances and impurities can be effectively removed, TritonX-100 loss of the tested object can be effectively prevented in the determination process, and the accuracy of a determination result is guaranteed. Meanwhile, the method provided by the invention is quick and convenient and can objectively evaluate the quality of a product.

Description

[0001] technical field [0002] The invention belongs to the field of biotechnology, and in particular relates to a method for determining the residual amount of Triton X-100 in antitoxin / antiserum. Background technique [0003] Antitoxin and antiserum (Antitoxin / Antiserum) refer to the use of specific antigens to immunize animals, then collect high-titer immune serum, and after extraction and purification, prepare complete antibodies (IgG) or antibody fragments [F(ab’) 2 ] immunoglobulin products. For the prevention and treatment of infections caused by the corresponding diseases. [0004] Because the raw materials of antitoxin and antiserum are derived from horse plasma, and horses are raised in an open environment, they may carry more viruses, so the products have biological safety problems. In order to ensure the safety of this product, in addition to controlling the breeding environment of immunized horses and the quality of horse plasma, how to effectively remove or ...

Claims

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Application Information

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IPC IPC(8): G01N30/89
Inventor 吴笛杨冬罗靖雄
Owner 玉溪九洲生物技术有限责任公司
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